JP2006193536A - Stable, aqueous alpha interferon solution formulation - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a re-formulation of an α-type interferon solution product which maintains, for an extended storage period, high chemical stability, high physical stability, and a high biological activity of an α-type interferon in an aqueous solution formulation, and is used for preparing a solution formulation which does not contain a human-blood-derived product such as HSA (human serum albumin). <P>SOLUTION: The formulation is a stable aqueous formulation having a high biological α-type interferon activity, does not contain a human-blood-derived product, and contains the followings: (a) an α-type interferon of 0.1×10<SP>6</SP>-100×10<SP>6</SP>IU/mL; (b) a buffer to maintain the pH in the range of 4.5-7.1; (c) a chelating agent in an effective quantity; (d) a sorbitan mono-9-octadecenoate poly(oxy-1,2-ethanediyl) derivative in a quantity sufficient to stabilize an α-type interferon against the loss of the α-type interferon; (e) a tonicity agent in an effective quantity; (f) an antimicrobial preservative in an effective quantity; and (g) the water for injection in a quantity sufficient for preparing a solution containing the ingredients listed above. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

BACKGROUND OF THE INVENTION The present invention is a stable, aqueous formulation that is free of human serum-derived products and maintains high biological activity and high chemical and physical stability of alpha interferon for extended periods of time. About.

  U.S. Pat.No. 4,496,537 discloses a biologically stable aqueous solution of alpha interferon containing alpha interferon, human serum albumin and alanine or glycine, water, and a buffer system to maintain the pH at 6.5-8.0. To do. Human serum albumin ("HSA") acts as a stabilizer for alpha interferon and is due to the coating and / or adsorption of alpha interferon on the stainless steel and glass surfaces of compounding containers, processing equipment and storage containers Prevent loss of alpha interferon from solution. Solution formulations containing alpha interferon and HSA can be used for chemical and biological stability of alpha interferon when such a solution is stored at 2-8 ° C for an extended period of time (ie, greater than 2 years). Maintaining sex.

  In recent years, the global AIDS epidemic has resulted in a health registration agency requiring manufacturers to be wary of products containing human blood-derived products such as HSA (eg, alpha interferon) It was.

  Solution formulations that do not contain human blood-derived products such as HSA while maintaining high chemical and physical stability and high biological activity of alpha interferon in aqueous formulations for extended storage periods There is a need to re-form the alpha interferon solution product in order to obtain

The present invention provides the following:
(1) A stable, aqueous formulation having high biological α-type interferon activity and free of human blood-derived products, comprising:
a. Alpha interferon of 0.1 × 10 ^{6 to} 100 × 10 ⁶ IU / mL;
b. a buffer system for maintaining the pH in the range of 4.5 to 7.1;
c. An effective amount of chelating agent;
d. a sufficient amount of sorbitan mono-9-octadecenoate poly (oxy-1,2-ethanediyl) derivative to stabilize the alpha interferon against loss of alpha interferon;
e. An effective amount of a tonicity agent;
f. An effective amount of an antimicrobial preservative; and g. A sufficient amount of water for injection to prepare a solution of the ingredients listed above,
A formulation containing
(2) The formulation according to item 1, wherein the buffer system is disodium phosphate and monosodium phosphate.
(3) The formulation according to item 1, wherein the chelating agent is edetate disodium or citric acid.
(4) A formulation according to item 1, wherein the tonicity agent is sodium chloride.
(5) The formulation according to item 1, wherein the preservative is selected from m-cresol, phenol, methylparaben, propylparaben, or a mixture thereof.
(6) The formulation according to item 1, wherein the α-type interferon is interferon α2.
(7) A stable aqueous formulation having high biological interferon α2 activity and free of human serum albumin, comprising:
mg / mL
a. Interferon α2 5 × 10 ⁶ to 50 × 10 ⁶ IU
b. Anhydrous disodium phosphate 1.8
c. Monosodium phosphate monohydrate 1.3
d. Ethylenediaminetetraacetic acid disodium 2 hydrogen 0.1
e. Sorbitan mono-9-octadecenoate poly (oxy-1,2-ethanediyl) derivative 0.1
f. Methylparaben 1.2
g. Propylparaben 0.12
h. Sodium chloride; and 7.5
i. Formulation containing water for injection, up to 1 mL.
(8. A stable aqueous formulation with high biological interferon α2 activity and no human serum albumin, comprising:
mg / mL
a. Interferon α2 5 × 10 ⁶ to 50 × 10 ⁶ IU
b. Anhydrous disodium phosphate 1.8
c. Monosodium phosphate monohydrate 1.3
d. Ethylenediaminetetraacetic acid disodium 2 hydrogen 0.1
e. Sorbitan mono-9-octadecenoate poly (oxy-1,2-ethanediyl) derivative 0.1
f. m-Cresol 1.5
g. Sodium chloride; and 7.5
h. Formulation containing water for injection, up to 1 mL.
(9) A product comprising a package and the formulation of any of items 1-8, wherein the package is a multi-dose glass vial.
(10) A product including a pre-filled syringe containing the formulation according to any one of items 1 to 8 in an effective amount.
(11) A product comprising a package and the formulation of any of items 1-8, wherein the package is a single dose vial.
(12) A process for preparing a stable, aqueous formulation having high biological α-type interferon activity and free of human blood-derived products, wherein the process converts an effective amount of α-type interferon to pH Make buffer systems, chelating agents, sorbitan mono-9-octadecenoate poly (oxy-1,2-ethanediyl) derivatives, tonicity agents, antimicrobial preservatives and aqueous solutions A process comprising mixing with sufficient water to do.
(13) A process according to item 12, wherein the aqueous solution is prepared and maintained substantially free of dissolved oxygen.
(Summary of the Invention)
The present invention is a stable, aqueous formulation that maintains high biological alpha interferon activity and is free of human blood-derived products, comprising:
a. Alpha interferon of 0.1 × 10 ^{6 to} 100 × 10 ⁶ IU / mL;
b. a buffer system for maintaining the pH in the range of 4.5 to 7.1;
c. An effective amount of chelating agent;
d. a sufficient amount of sorbitan mono-9-octadecenoate poly (oxy-1,2-ethanediyl) derivative to stabilize the alpha interferon against loss of alpha interferon;
e. An effective amount of a tonicity agent;
f. An effective amount of an antimicrobial preservative; and g. A sufficient amount of water for injection to prepare a solution of the ingredients listed above,
A formulation containing is provided.

The present invention is a stable, aqueous formulation having high alpha interferon biological activity and free of human blood derived products, comprising:
a. Alpha interferon of 0.1 × 10 ^{6 to} 100 × 10 ⁶ IU / mL;
b. Sufficient buffer system to maintain the pH of the solution in the range of 4.5 to 7.1;
c. About 0.01-1 mg / mL ethylenediaminetetraacetic acid disodium dihydrogen;
d. About 0.01-1 mg / mL sorbitan mono-9-octadecenoate poly (oxy-1,2-ethanediyl) derivative;
e. About 1-9 mg / mL sodium chloride;
f. An effective amount of an antimicrobial preservative selected from m-cresol, phenol, methylparaben, propylparaben, or mixtures thereof; and g. Water for injection to make 1 mL,
A formulation containing is provided.

In a preferred aspect, the present invention is a stable, aqueous formulation having high biological alpha interferon activity and free of human blood derived products, comprising:
mg / mL
a. α2 interferon 5 × 10 ⁶ to 50 × 10 ⁶ IU
b. Anhydrous disodium phosphate 1.8
c. Monosodium phosphate monohydrate 1.3
d. Ethylenediaminetetraacetic acid disodium 2 hydrogen 0.1
e. Polysorbate 80 0.1
f. Methylparaben 1.2
g. Propylparaben 0.12
h. Sodium chloride; and 7.5
i. Formulations containing up to 1 mL of water for injection are provided.

In another preferred aspect, the present invention further provides a stable, aqueous formulation having high biological alpha interferon activity and free of human blood derived products, comprising:
mg / mL
a. α2 interferon 5 × 10 ⁶ to 50 × 10 ⁶ IU
b. Anhydrous disodium phosphate 1.8
c. Monosodium phosphate monohydrate 1.3
d. Ethylenediaminetetraacetic acid disodium 2 hydrogen 0.1
e. Polysorbate 80 0.1
f. m-Cresol 1.5
g. Sodium chloride; and 7.5
h. Formulations containing up to 1 mL of water for injection are provided.

  The present invention also provides a process for the preparation of a stable, aqueous formulation having high biological alpha interferon activity and free of human blood derived products, the process comprising an effective amount of alpha interferon. Buffer systems, chelating agents, sorbitan mono-9-octadecenoate poly (oxy-1,2-ethanediyl) derivatives, tonicity agents, antimicrobial preservatives and solutions that can maintain pH within the range of 4.5-7.1 A process is provided that includes mixing with sufficient water to form. In a preferred aspect of the process of the present invention, the solution is prepared and maintained substantially free of dissolved oxygen, and the inert atmosphere headspace above the solution is less than about 4% (volume) oxygen. Maintained at the value of.

DETAILED DESCRIPTION Although the inventors do not contain human serum albumin, upon storage in the period 2 to 8 ° C. which extends for at least 24 months, high chemical for α interferon, biological and physical stability A specific amount of a specific set of ingredients was chosen that made it possible to develop an aqueous alpha interferon solution formulation that preserved the sex.

  The term “free from human blood-derived products” as used herein with respect to the formulations of the present invention means that human blood-derived products such as HSA are not used in the preparation of the solution formulations of the present invention.

The term “high chemical stability” as used herein with respect to α-type interferon in the formulations of the present invention means that α-type interferon has a chemical integrity of at least 85%, preferably 85% to 100%. , Means maintaining at 2-8 ° C. during storage for at least 24 months. See Tables 1 and 2. Chemical integrity is disclosed, for example, by TLNagabhushan et al., “Characterization of engineered α2 interferon” pages 79-88, Interferon Research Clinical Application and Regulatory Consideration , edited by Zoon et al., Elsevier Science Publishing Co., Inc. 1984. As determined by measurement of protein content in such HPLC assays (see results in Tables 1-4).

The term “high biological stability” as used herein with respect to α-type interferon in the formulation of the present invention refers to the fact that α-type interferon in the formulation is WPProtzman et al., J. Clinical Microbiology , (1985), 22 , 596-599, as measured in standard methods of inhibition of viral cytopathic effect (CPE), at least 75%, preferably at least 85%, more preferably 90% -100% Means maintaining its biological integrity upon storage at 2-8 ° C. for at least 24 months (see results in Tables 1-4).

  The term “high physical stability” as used herein with respect to α-type interferon in the formulations of the present invention remains clear upon storage of the present formulations at 2-8 ° C. for at least 24 months. I.e. no turbidity or visible particulate matter (i.e. particles larger than about 60-70 microns in diameter). See Tables 1, 2, and 3. The results listed in Tables 1, 2, and 3 show that most solution formulations containing protein products such as alpha interferon are visually observable upon extended storage at 2 ° C to 8 ° C. It is surprising in that it develops particles (i.e. particles larger than 60-70 microns in diameter). Test methods used to measure particulate matter in the solution formulations of the present invention (see Tables 1 to 4) are described in USP 23 / NF 18, United States Pharmacopeial Convention, Inc., (1995), Rockville, Maryland (see Physical Test <788> on pages 1813-1816). The method used to measure the visual description of the solution formulations of the present invention is also described in USP 23, “General Requirement Test and Assays <1> Injections”, pages 1650-1652.

The inventors have found that visible particles can be avoided by adding chelating agents to the formulations of the present invention. Exemplary suitable chelating agents include ethylenediaminetetraacetic acid disodium dihydrogen (EDTA or edetate disodium) or citric acid. The use of edetate disodium is preferred. Although we wish to be not bound by any theory, edetate disodium is effectively a trace metal cation (eg, Zn ²⁺ , Fe ²⁺ , Cu ²⁺ or Al ³⁺ ) (these It is believed that the ions are complexed with excipients and packaging components (such as may be present in rubber stoppers or gaskets). Because edetate disodium has a higher affinity for these metal cations than alpha interferon, metal cations and alpha interferon that cause insoluble complex formation (eg, in the form of visible particles) and loss of activity. Interaction with is avoided. Effective amounts of chelating agents range from 0.01 to 1 mg / mL based on 0.1 × 10 ^{6 to} 100 × 10 ⁶ alpha interferon international units (“IU”) / mL. Preferably, 0.1 mg of edetate disodium is used for 5 × 10 ⁶ to 50 × 10 ⁶ IU α2 interferon.

Suitable buffer systems for the formulations of the present invention are those that maintain the pH of the aqueous formulation in the range of 4.5 to 7.1, preferably in the range of 6.5 to 7.1, and most preferably 6.8. The use of a disodium phosphate and monosodium phosphate buffer system is preferred. Typically, a preferred monosodium phosphate / disodium phosphate buffered 0.005-0.1 molar buffer is used for formulations containing 0.1 × 10 ^{6 to} 100 × 10 ⁶ IU alpha interferon per mL. The Other suitable buffer systems for maintaining the desired pH range of 4.5 to 7.1 include sodium citrate / citric acid and sodium acetate / acetic acid.

  A tonicity agent useful in the present invention is any agent that can impart an osmotic pressure equal to that of human serum to a formulation of the present invention. Exemplary suitable tonicity agents include sodium chloride, mannitol, glycine, glucose, and sorbitol. The use of sodium chloride as a tonicity agent is preferred.

When the formulation of the present invention contains 0.1 × 10 ^{6 to} 100 × 10 ⁶ IU α-type interferon per mL, the amount of tonicity agent used ranges from 1 to 10 mg / mL. For 5 × 10 ⁶ to 50 × 10 ⁶ IU α-type interferon per mL in the formulation of the present invention, the use of 7.5 mg / mL sodium chloride is preferred.

Injectable formulation of sorbitan mono-9-octadecenoate poly (oxy-1,2-ethanediyl) derivative, such as polysorbate 80 or polysorbate 20, containing alpha interferon of alpha interferon protein such as alpha 2b interferon It is useful as a stabilizer to prevent adsorption of stainless steel and glass surfaces of equipment used to make indictable formations. Effective amounts of polysorbate 20 or polysorbate 80 in the formulations of the present invention range from 0.01 to 1.0 mg / mL for formulations containing 0.1 × 10 ^{6 to} 100 × 10 ⁶ IU alpha interferon per mL. The use of polysorbate 80 is preferred. The use of 0.1 mg / mL polysorbate 80 is more preferred in all solution formulations of the present invention. If the concentration of α-type interferon such as α2 interferon is less than about 15 × 10 ⁶ IU / mL (eg 6 × 10 ⁶ IU / mL), it is due to adsorption of α-interferon in the absence of polysorbate 80 This loss of activity significantly reduces the biological activity of the formulation. Surprisingly, the inventors have found that polysorbate 80 prevents loss of α2b interferon and allows systemic delivery of α2b interferon without loss of biological activity. In the course of developing the formulations of the present invention, we have surprisingly found that polysorbate 80 has superior chemical properties compared to other nonionic surfactants (eg, Pluronic F127 and Pluronic F-68). And found to provide biological stability to α2b interferon.

The amount of alpha interferon useful in the formulations of the present invention is in the range of 0.1 × 10 ^{6 to} 100 × 10 ⁶ IU / mL, preferably in the range of 5 × 10 ⁶ to 50 × 10 ⁶ IU / mL.

  As used herein, the term “alpha interferon” refers to a family of highly homologous species-specific proteins that inhibit viral replication and cell growth and modulate the immune response. Representative suitable alpha interferons are interferons α2a such as ROFERON A interferon α2a available from Hoffmann-La Roche, Nutley, NJ, and interferons α2b such as INTRON A interferon α2b available from Schering Corporation, Kenilworth, NJ. Interferon α2c, such as BEROFOR interferon α2c available from Boehringer Ingelheim Pharmaceutical, Inc., Ridgefield, CT, available as SUMIFERON from Sumitomo, Japan, or as WELLFERON from The Wellcome Foundation Ltd., London, Great Britain Interferon αn1 (a purified mixture of natural α-interferons), or the consensus α-interferon available from Amgen, Inc., Newbury Park, California, or manufactured by Interferon Sciences and from ALFERON from PurdueFrederick Co., Norwalk, CT. Interface available under the trade name Ron αn3 (mixture of natural α interferons) is included. The use of interferon α2a or α2b is preferred. The use of interferon α2b is more preferred.

Antimicrobial preservatives found to be useful in the present invention include m-cresol, phenol, and methyl-paraben and propylparaben, and mixtures of the preservatives listed above (eg, phenol-methylparaben mixtures). To do. Effective amounts of m-cresol found to be useful in the present invention range from 0.5 to 2 mg / mL for formulations containing 0.1 × 10 ^{6 to} 100 × 10 ⁶ IU / mL alpha interferon . For formulations containing 5 × 10 ⁶ to 50 × 10 ⁶ IU / mL interferon α2b, the use of 1.5 mg / mL m-cresol is preferred.

Effective amounts of phenol found to be useful range from 0.5 to 5 mg / mL for solution formulations containing 0.1 × 10 ^{6 to} 100 × 10 ⁶ IU / mL α-type interferon.

When the formulation of the present invention contains 0.1 × 10 ^{6 to} 100 × 10 ⁶ IU / mL α-type interferon, the effective amount of methylparaben is in the range of 0.6 to 1.8 mg / mL, and the effective amount of propylparaben is The range is from 0.06 to 0.18 mg / mL.

When the formulation of the present invention contains 0.1 × 10 ^{6 to} 100 × 10 ⁶ IU / mL α2b interferon, it is preferred to use 1.2 mg / mL methyl paraben in combination with 0.12 mg / mL propyl paraben.

  More preferred is the use of m-cresol as an antimicrobial preservative.

  The water used for the preparation of the formulation of the present invention is preferably water for injection.

  Development of the aqueous formulation of the present invention that maintains the high biological activity and high chemical and physical stability of alpha interferon over extended storage periods without using HSA as a stabilizer. In the process, we have developed a sorbitan mono-9-octadecenoate poly (oxy-1,2-ethanediyl such as polysorbate 80 that is required to act as a stabilizer for alpha interferon. ) The amount of derivative in the aqueous formulation to provide adequate antimicrobial protection to the formulation in accordance with various global health registration requirements without causing the formation of unwanted turbidity in the solution. It was identified that it had a direct effect on the effective amount of antimicrobial preservative that could be added.

  Therefore, when the preferred stabilizer polysorbate 80 is present in the formulation of the present invention (at a preferred effective amount of 0.1 mg / mL), a preferred antimicrobial that can be added without causing cloudiness of the formulation An effective amount of preservative (eg, m-cresol) has been found to be exact. For example, turbidity is observed when the amount of m-cresol added to a formulation containing 0.1 mg / mL polysorbate 80 (as shown in Example 3) increases to greater than 1.75 mg / mL. It was. Similar turbidity problems were observed when the amount of polysorbate 80 in the resulting formulation varied from 0.01 mg / mL to 1 mg / mL. 1.75 mg / mL or less, preferably about 1.5 mg / mL m-cresol, when added to a formulation prepared according to the procedure of Example 3 (which contains 0.1 mg / mL polysorbate 80) Was not observed. This stringency was also observed for parabens and phenols when they were used as antimicrobial preservatives. For formulations of the invention containing 0.01-1 mg / mL polysorbate 80, the effective amount of methyl paraben to avoid turbidity when used with 0.12 mg / mL propylparaben is about 1.2 mg / mL or less And the effective amount of phenol to avoid turbidity (when phenol is used instead of parabens) should range from 0.5 to less than about 4 mg / mL.

  Alpha interferon formulations are useful for the treatment of various medical conditions such as renal cell carcinoma, AIDS-related Kaposi's sarcoma, chronic and acute hepatitis B, chronic and acute non-A, non-B / C hepatitis. The formulations of the present invention are useful in treating these conditions, preferably as injectable aqueous solutions.

Examples The following non-limiting examples illustrate the preparation of aqueous solutions of alpha interferon.

The procedure listed after Example 5 is used to prepare the inventive formulations of Examples 1-5.
Example 1
Active substance: Interferon α2b 0.1 × 10 ^{6 to} 100 × 10 ⁶ IU / mL ^*
Buffer solution: Sodium phosphate 0.005-0.1M
(1 sodium / 2 sodium)
Chelating agent: edetate disodium 0.01-1mg / mL
Stabilizer: Polysorbate 80 0.01-1mg / mL
Tonicity adjusting agent: sodium chloride 1-9mg / mL
Antimicrobial preservative: m-cresol 0.5-1.75mg / mL
Or phenol 0.5- <4mg / mL
Or methylparaben 0.6-1.2mg / mL
Propylparaben 0.06-0.12mg / mL
Solvent: Water for injection up to 1mL
^* IU-International units
Example 2
Interferon α2b 10 × 10 ⁶ IU / mL
Anhydrous disodium phosphate 1.8mg / mL
Monosodium phosphate monohydrate 1.3mg / mL
Edetate disodium 0.1mg / mL
Polysorbate 80 0.1mg / mL
Methylparaben 1.2mg / mL
Propylparaben 0.12mg / mL
Sodium chloride 7.5mg / mL
Water for injection up to 1mL
Example 3
Interferon α2b 10 × 10 ⁶ IU / mL
Anhydrous disodium phosphate 1.8mg / mL
Monosodium phosphate monohydrate 1.3mg / mL
Edetate disodium 0.1mg / mL
Polysorbate 80 0.1mg / mL
m-cresol 1.5mg / mL
Sodium chloride 7.5mg / mL
Water for injection up to 1 mL Stability data for Examples 2 and 3 are summarized in Tables 1 and 2, respectively.

Example 4
The formulation of Example 3 was prepared using 6 × 10 ⁶ IU / mL α2b interferon according to the manufacturing method detailed herein below, using a nitrogen sparging of the solution, and about 4 oxygen in the headspace. % (Volume) or less.

  Vials containing the indicated volume of 3 mL of solution were stored at 30 ° C, 25 ° C and 4 ° C. The results are summarized in Table 3.

Example 5
The formulation of Example 4 was used in all respects except that the solution was not sparged or overlaid with nitrogen and the headspace oxygen volume was about 20% (volume) as found in ambient air. It was prepared according to the production method described in detail below.

  Vials containing the indicated volume of 3 mL of solution were stored at 30 ° C, 25 ° C and 4 ° C. The results are summarized in Table 4.

  Similar results are expected when interferon α2b in Examples 1-5 is replaced with equivalent amounts of Roferon A, Wellferon or Sumiferon interferon α.

Production Method A to Example 1-5 Formulation of paraben-containing aqueous formulation as shown in Example 2 Approximately 80% of water for injection is placed at a temperature above 70 ° C. into a jacketed container equipped with a stirrer.

    2. Separately, about 30% water for injection is placed in another suitable container. Cool and maintain water temperature between 20 ° C and 25 ° C. Begin sparging with filtered nitrogen and stratification into the water used to bring the batch to final volume to maintain the dissolved oxygen level below 0.25 ppm.

    3. Into the compounding vessel of Step 1, add metipparaben and propylparaben and dissolve with mixing while maintaining the temperature of the solution between 70 ° C and 80 ° C.

    4). Cool the solution of step 3 to a temperature between 20 ° C and 25 ° C. Sprinkle and overlay the solution with filtered nitrogen. Maintain dissolved oxygen level below 0.25 ppm.

5. Add the following ingredients to the solution of step 4 and dissolve with mixing while maintaining nitrogen sparging and stratification:
Anhydrous disodium phosphate monosodium phosphate monohydrate edetate disodium sodium chloride Stop nitrogen sparging of the solution in step 5. Maintain a nitrogen overlay in the compounding vessel.

    7). Polysorbate 80 is placed in about 50 mL of water for injection (for a 1 liter size batch) in a separate container and dissolved. Transfer the polysorbate 80 solution to the solution in step 6.

    8). Check the pH of the solution. The pH should be between 6.6 and 7.0. pH adjustment is not necessary.

    9. Place the interferon α2b bulk drug solution into the solution of step 8 with mixing.

    Ten. Water for injection that has been sparged with nitrogen (from step 2) is added to bring the batch to the final volume. Gently stir the solution until homogeneous.

    11． Aseptically, the solution is washed and filtered through a sterile filter that has been tested for integrity. The sterilized solution is collected in a sterilized filled container layered with sterile filtered nitrogen. The filter is integrity tested after filtration.

    12． Step 11 filling container is overlaid with sterile filtered nitrogen and sealed.

B. The manufacturing procedure used to prepare an aqueous solution containing m-cresol (as shown in Example 3) as a formulation preservative for an m-cresol-containing aqueous formulation as shown in Example 3 is Exactly the same as above in this specification, except that the temperature is maintained between 20 ° C. and 25 ° C. and m-cresol is added after step 6.

C. Formulating an HSA-free aqueous alpha interferon solution formulation under ambient air All steps are performed under ambient air, nitrogen is not sparged or overlaid on the solution, and ambient air (usually about 20% (volume) oxygen As the formulation of Example 5 using the manufacturing procedure used to prepare the HSA-free aqueous alpha interferon formulation of Examples 1-4. A simple formulation was prepared.

  In order to maintain high chemical, physical and biological stability, the water used to prepare the aqueous alpha interferon solution and the aqueous alpha interferon solution so formed are substantially free of dissolved oxygen. Preferably, the aqueous solution is made and stored with an inert atmosphere (eg, nitrogen) headspace containing no more than about 4% (volume) oxygen. The term “substantially free of dissolved oxygen” as used herein means an oxygen level of about 0.25 ppm or less at a water temperature of about 20 ° C. to 25 ° C. Typically, this preferred dissolved oxygen level of 0.25 ppm is sufficient for an inert atmosphere (eg, nitrogen gas) to the water used to prepare the aqueous solution (temperature maintained at about 20 ° C. to 25 ° C.). Conveniently achieved by spraying for a time (eg, about 30 minutes) to reduce the dissolved oxygen to a value of about 0.25 ppm or less. Spraying is continued throughout the manufacturing procedure to maintain the dissolved oxygen level at 0.25 ppm. We have an aqueous formulation of the invention having a dissolved oxygen level of 1 ppm and an oxygen content of 7% (volume) headspace stored under the same conditions after 3 months storage at 25 ° C. It was found that it showed significantly greater loss of alpha interferon chemical stability compared to a similar aqueous formulation having a preferred dissolved oxygen level of 0.25 ppm and an oxygen content of 4% (volume) headspace. .

  By side-by-side comparison of the α2b interferon solution stability data shown in Tables 3 and 4, the aqueous solution formulations of the present invention prepared under the nitrogen / hypoxic conditions used in Example 4 It is shown that there is no significant stability difference over 12 months at 4 ° C. with the formulation prepared under atmospheric air according to Example 5. In contrast, a comparison of α2b interferon stability in the solutions of Examples 4 and 5 stored at higher temperatures (eg, 25 ° C. and 30 ° C.) simultaneously reduced the oxygen content in the headspace to about 4% (vol.) The protective effect achieved by the preferred (safer) use of nitrogen sparging, which results in low dissolved oxygen levels in the aqueous solution while maintaining the value of.

  The aqueous formulation of the present invention may be stored in any suitable washed and sterilized filled container or container (eg, a 2-mL or 5-mL Type I flint glass vial stoppered with a gray butyl rubber stopper). . The aqueous solution formulations of the present invention can also be stored in a prefilled multi-dose syringe (eg, a syringe useful for the delivery of an indictable solution of a drug such as insulin). . A typical suitable syringe includes a system that includes a pre-filled vial attached to a pen-type syringe (eg, Novolet Novo Pen available from NovoNordisk). Exemplary suitable systems include pre-filled, pen-type syringes that allow easy self-injection and accurate and reproducible dose setting by the user.

  The aqueous formulation of the present invention (eg, as shown in the examples) can also be lyophilized to form a powder for reconstitution. The lyophilized alpha interferon powder is expected to maintain its chemical and biological stability for at least 2 years when stored at 2-8 ° C.

Claims (1)

Process described in the examples.